Search

Denosumab is the world's first human monoclonal antibody for the treatment of osteoporosis and shares an active pathway with bisphosphonates, strongly suppressing osteoclast activities. This is the first case report describing a possible relationship between the development of osteonecrosis of external auditory canal and denosumab administration. We herein report an 81-year-old woman diagnosed with left osteonecrosis of external auditory canal who had a history of denosumab administration. She underwent left radical mastoidectomy due to being refractory to conservative treatment. No major complications or recurrence were observed in the left ear after surgery, but bone erosion in the right ear has continued to progress slowly despite the cessation of denosumab administration. Otolaryngologists should be aware of the association between osteonecrosis of external auditory canal and denosumab administration and consider performing long-term observation even after cessation., (Copyright © 2020. Published by Elsevier B.V.)

Objective: The recent development of extended reality technology has attracted interest in medicine. We explored the use of patient-specific virtual reality (VR) and mixed reality (MR) temporal bone models in anatomical teaching, pre-operative surgical planning and intra-operative surgical referencing., Methods: VR and MR temporal bone models were created and visualized on head-mounted display (HMD) and MR headset respectively, by a novel webservice that allows users to convert computed tomography images to VR and MR images without specific knowledge of programming. Eleven otorhinolaryngology trainees and specialists were asked to manipulate the healthy VR temporal bone model and to assess its validity by filling out a questionnaire. Additionally, VR and MR pathological models of petrous apex cholesteatoma were utilized for surgical planning pre-operatively and for referring to the anatomy during the surgery., Results: Most participants were favorable about the VR model and considered HMD as superior to a flat computer screen. 91% of the participants agreed or somewhat agreed that VR through HMD is cost effective. In addition, the VR pathological model was used for planning and sharing the surgical approach during a pre-operative surgical conference. The MR headset was worn intra-operatively to clarify the relationship between the pathological lesion and vital anatomical structures., Conclusion: Regardless of the participants' training level in otorhinolaryngology or VR experience, all participants agreed that the VR temporal bone model is useful for anatomical education. Furthermore, the creation of patient-specific VR and MR models using the webservice and their pre- and intra-operative usages indicated the potential of innovative adjunctive surgical instrument., Competing Interests: Declaration of Competing Interest Maki Sugimoto is a board member of Holoeyes Inc., (Copyright © 2021. Published by Elsevier B.V.)

Objectives: We aim to clarify the frequency of lymph node metastasis of external auditory canal (EAC) carcinoma, including susceptible locations, adequate extent of elective neck dissection, and the relationship between the tumor infiltration site and lymph node metastasis., Patients and Methods: From 2003 to 2018, 63 patients with EAC carcinoma at Tokyo Medical and Dental University Hospital were enrolled in this study. The T and N stages, locations of clinically positive lymph nodes, prognoses, and anatomic site of tumor infiltration were analyzed after treatment., Results: Clinically positive lymph node metastasis (cN+) was detected in 18 patients (28.6%), consisting of T1, T2, T3, and T4 disease in 1 (6%), 2 (22%), 8 (38%), and 7 (41%) patients, respectively. The metastatic locations were at level II in 10 patients, parotid gland nodes in 7, preauricular nodes in 5, level Ib in 3, level Va in 3, level III in 1, and superficial cervical nodes in 1. Neck recurrence was determined in two of 45 patients with clinically negative lymph nodes (cN0), with the metastatic locations being levels II, Ib, and III. Among 18 cN+ cases, neck recurrence was noted in 2 of 9 patients who underwent neck dissection. Neck lesions were found to be manageable in all five patients who underwent docetaxel, cisplatin, 5-fluorouracil, and radiation therapy (TPF-RT). No relationship was noted between the tumor infiltration site and lymph node metastasis among T3/4 canrcinoma patients., Conclusions: Elective neck dissection could be indicated only in T3/4 patients with free flap reconstruction. Levels Ib to III are considered appropriate for elective neck dissection in cN0 cases. Levels Ib to III and Va indicated favorable sites, even in cases with metastasis in the parotid gland or preauricular area. Furthermore, TPF-RT could be a useful option even in cN+ cases., Competing Interests: Declaration of Competing Interest There are no conflicts of interest to declare., (Copyright © 2020. Published by Elsevier B.V.)

Polyunsaturated fatty acids (PUFAs), represented by the omega-6 fatty acid arachidonic acid (AA) and omega-3 fatty acid docosahexaenoic acid (DHA), are essential components of the human body. PUFAs are converted enzymatically into bioactive lipid mediators, including AA-derived cysteinyl leukotrienes (cys-LTs) and lipoxins and DHA-derived protectins, which orchestrate a wide range of immunological responses. For instance, eosinophils possess the biosynthetic capacity of various lipid mediators through multiple enzymes, including 5-lipoxygenase and 15-lipoxygenase, and play central roles in the regulation of allergic diseases. Dysregulated metabolism of PUFAs is reported, especially in severe asthma, aspirin-exacerbated respiratory disease, and eosinophilic chronic rhinosinusitis (ECRS), which is characterized by the overproduction of cys-LTs and impaired synthesis of pro-resolving mediators. Recently, by performing a multi-omics analysis (lipidomics, proteomics, and transcriptomics), we demonstrated the metabolic derangement of eosinophils in inflamed tissues of patients with ECRS. This abnormality occurred subsequent to altered enzyme expression of gamma-glutamyl transferase-5. In this review, we summarize the previous findings of dysregulated PUFA metabolism in allergic diseases, and discuss future prospective therapeutic strategies for correcting this imbalance., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Purpose: To investigate flow signal within subretinal hyperreflective material (SHRM) using optical coherence tomography angiography (OCTA) and its association with aflibercept treatment responses in treatment-naïve neovascular age-related macular degeneration (nAMD)., Design: Prospective consecutive interventional case series., Methods: Forty-four eyes of 44 patients with treatment-naïve nAMD manifesting SHRM on OCT were studied. All patients underwent OCTA and received 3 monthly aflibercept injections. The intrinsic flow signals within SHRM were quantitatively analyzed using OCTA, and eyes were classified into the vascular and avascular SHRM groups., Results: Of 44 eyes, 21 (47.7%) and 23 (52.3%) showed vascular SHRM and avascular SHRM, respectively. Compared with eyes with avascular SHRM, eyes with vascular SHRM showed higher rates of external limiting membrane (ELM) disruption owing to SHRM (P = .015), classic choroidal neovascularization (CNV) (85.7% vs 26.1%, P = .87 × 10 -4 ), and intraretinal fluid (P = .008) at baseline. After 3 aflibercept injections, 38 eyes (86.4%) showed dry macula despite persistent SHRM in 24 eyes (54.5%). Compared with the eyes with resolved SHRM, those with persistent SHRM showed higher rate of vascular SHRM (75.0% vs 15.0%, P = .86 × 10 -4 ), classic CNV (P = .032), absence of polypoidal lesion (P = .020), ELM disruption owing to SHRM (P = .042), and intraretinal fluid (P = .008). Dry macula after loading injections was significantly associated with SHRM resolution (P = .025)., Conclusions: In nAMD, SHRM can be categorized as vascular and avascular by quantitative OCTA analysis. Vascular SHRM persisted after treatment and was associated with failure to achieve dry macula, suggesting that vascular SHRM is predictive of lower response to anti-VEGF therapy., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Objective: To assess possible delayed recovery of the maximum speech discrimination score (SDS) when the audiometric threshold ceases to change., Methods: We retrospectively examined 20 patients with idiopathic sudden sensorineural hearing loss (ISSNHL) (gender: 9 males and 11 females, age: 24-71 years). The findings of pure-tone average (PTA), maximum SDS, auditory brainstem responses (ABRs), and tinnitus handicap inventory (THI) were compared among the three periods of 1-3 months, 6-8 months, and 11-13 months after ISSNHL onset., Results: No significant differences were noted in PTA, whereas an increase of greater than or equal to 10% in maximum SDS was recognized in 9 patients (45%) from the period of 1-3 months to the period of 11-13 months. Four of the 9 patients showed 20% or more recovery of maximum SDS. No significant differences were observed in the interpeak latency difference between waves I and V and the interaural latency difference of wave V in ABRs, whereas an improvement in the THI grade was recognized in 11 patients (55%) from the period of 1-3 months to the period of 11-13 months., Conclusion: The present study suggested the incidence of maximum SDS restoration over 1 year after ISSNHL onset. These findings may be because of the effects of auditory plasticity via the central auditory pathway., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Objective: To characterize the long-term outcomes of meatoplasty using our new technique, inferiorly based retroauricular island pedicle flap for external auditory canal (EAC) stenosis., Methods: A long-term clinical analysis of meatoplasty for nine patients (mean age, 33 years; age range, 8-64 years) with stenotic EAC was performed. The follow-up period after meatoplasty ranged from 5 years to 14 years, with a mean duration of 8.4 years. We compared preoperative and postoperative otoscopic findings, high-resolution computed tomographic (CT) images of the EAC, and hearing levels., Results: Otoscopic examinations demonstrated widened EAC in eight of the nine patients. There was a significant increase in the size of the EAC on postoperative CT examinations when compared with the preoperative state. The hearing outcome in all patients was variable. The postoperative air-bone gap (ABG) was closed to 10dB or less in two cases, while the other seven cases showed ABG ranging from 15.0dB to 57.5dB., Conclusions: We performed meatoplasty using inferiorly based retroauricular island pedicle flap for nine patients with EAC stenosis and eight of the nine patients demonstrated satisfactory patent EAC during a mean follow-up of 8.4 years., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Purpose: To investigate whether the photoreceptor layer status of the detached retina, which has the potential to plug the macular hole (MH), can predict postoperative visual acuity (VA)., Design: Retrospective, consecutive, observational case series., Methods: A consecutive series of 48 eyes in 48 patients with surgically closed MHs were recruited into the study. Spectral-domain optical coherence tomography (SD OCT) images were obtained preoperatively and postoperatively. Length gaps between photoreceptor layers and the preoperative diameter of the basal MH in the detached part of the retina was calculated for the photoreceptor outer segment (OS) and the external limiting membrane (ELM). The postoperative photoreceptor layer status at the fovea, including continuity of the ELM line, inner segment ellipsoid line (ISe), and cone outer segment tip (COST) line, was categorized by graders., Results: The preoperative OS gap length and ELM gap length correlated significantly with postoperative VA. A larger OS or ELM gap length was associated with a more severe postoperative photoreceptor layer status. Eyes with a moderately reflective lesion had a significantly longer OS gap length and ELM gap length, significantly worse postoperative VA, and more severe photoreceptor layer status than did eyes without such a moderately reflective lesion. Eyes with a better postoperative COST status had significantly better postoperative VA (P = .0465)., Conclusions: OS and ELM gap length, which represent the degree of insufficiency of MH plugging, was strongly correlated with postoperative VA; moreover, these preoperative parameters were associated with the postoperative photoreceptor layer status, including glial cell proliferation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Sertraline is one of the serotonin-specific reuptake inhibitors that is effective in treating several disorders such as major depression, obsessive-compulsive disorder, panic disorder, and social phobia. It is marketed in the form of its hydrochloride salt, which exhibits better solubility in water than its free base form. However, the absorption of sertraline through biological membranes could be improved by enhancing the solubility of its base because it is more hydrophobic than sertraline hydrochloride. To clarify the mechanism for the interaction of sertraline base with β-CD, it is important to study the basic interaction between the β-CD ring and sertraline base. Therefore, in this study, the currently used hydrochloride salt form was converted into the free base and β-CD was used as a model for β-CD derivatives to evaluate the interaction between β-CD and the sertraline base. The solid-state physicochemical characteristics of the sertraline-β-CD complex were investigated by the phase solubility method, differential scanning calorimetry, Fourier transform IR spectroscopy, FT-Raman spectroscopy, powder X-ray diffraction, and (13)C cross-polarization magic-angle spinning NMR measurements. The results showed that sertraline base and β-CD form an inclusion complex, and the stoichiometric ratio of the solid-state sertraline base-β-CD complex is 1:1, which was estimated by the (1)H NMR measurements of the complex dissolved in DMSO-d6., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objective: Mutations of transmembrane channel-like 1 gene (TMC1) can cause dominant (DFNA36) or recessive (DFNB7/B11) deafness. In this article, we describe the characteristics of DFNA36 and DFNB7/B11 deafness, the features of the Tmc1 mutant mouse strains, and recent advances in our understanding of TMC1 function., Methods: Publications related to TMC1, DFNA36, or DFNB7/B11 were identified through PubMed., Results: All affected DFNA36 subjects showed post-lingual, progressive, sensorineural hearing loss (HL), initially affecting high frequencies. In contrast, almost all affected DFNB7/B11 subjects demonstrated congenital or prelingual severe to profound sensorineural HL. The mouse Tmc1 gene also has dominant and recessive mutant alleles that cause HL in mutant strains, including Beethoven, deafness, and Tmc1 knockout mice. These mutant mice have been instrumental for revealing that Tmc1 and its closely related paralog Tmc2 are expressed in cochlear and vestibular hair cells, and are required for hair cell mechanoelectrical transduction (MET). Recent studies suggest that TMC1 and TMC2 may be components of the long-sought hair cell MET channel., Conclusion: TMC1 mutations disrupt hair cell MET., (Published by Elsevier Ireland Ltd.)

The objective of this study is to evaluate the effect of polymeric nanoparticles (NPs) on the allergic response of mast cells that release inflammatory mediators such as histamine through exocytosis. Submicron-sized biodegradable poly(DL-lactide-co-glycolide) (PLGA) NPs were prepared by the emulsion solvent diffusion method. Here, we examined the interactions of the mast cells with two types of PLGA NPs, unmodified NPs and NPs modified with chitosan (CS), a biodegradable cationic polymer. The cellular uptake of NPs increased by CS modification due to electrostatic interactions with the plasma membrane. NPs were taken up by mast cells through an endocytic pathway (endocytic phase) and then the cellular uptake was saturated and maintained plateau level by the exclusion of NPs through exocytosis (exocytic phase). Antigen-induced histamine release from mast cells was inhibited during the exocytic phase. The extent of histamine release inhibition was related to the amount of excluded NPs. Exocytic exclusion of NPs competitively antagonize the antigen-induced exocytotic release of histamine by highjacking exocytosis machinery such as SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins, since histamine release was recovered in mast cells that overexpress SNAP-23. The inhibitory effect of the allergic response by PLGA NPs was also evaluated in vivo using the mouse model for systemic anaphylaxis. The administration of NPs suppressed the antigen-induced systemic allergic response in vivo. In conclusion, PLGA NP itself has actions to inhibit the allergic responses mediated by mast cells., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

In this study, we investigated surface-modified nanoparticles (NP) formulated using a biodegradable polymer, poly(D,L-lactide-co-glycolide) (PLGA), for targeting central nervous system (CNS) diseases. Polysorbate 80 (P80), poloxamer 188 (P188), and chitosan (CS) were used to modify the surfaces of PLGA NP to improve the brain delivery of NP. Surface-modified PLGA NP were formulated using an emulsion solvent diffusion method. 6-Coumarin was used as a fluorescent label for NP. The different formulations of 6-coumarin-loaded PLGA NP were injected into rats via carotid arteries. NP remaining in the brain were evaluated quantitatively, and brain slices were observed using confocal laser scanning microscopy (CLSM). Carotid artery administration was more effective for delivering NP into the brain compared to intravenous administration. After administration, NP concentrations in the brain were increased by NP surface modification, especially CS- and P80-PLGA NP. CLSM observations indicated that P80-PLGA NP could cross the blood-brain barrier and thus serve as a drug delivery system for the CNS. These results indicate that surface-modified PLGA NP have a high potential for use in CNS delivery systems., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Chitosan (CS)-modified poly(D,L-lactide-co-glycolide) (PLGA) nanospheres (NS) were developed and evaluated for use with a nuclear factor kappa B (NF-κB) decoy oligonucleotide (ODN) oral delivery system in an experimental model of ulcerative colitis (UC). Decoy ODN-loaded PLGA NS were prepared by an emulsion solvent diffusion method, and the physicochemical properties of NS were investigated. CS-modified PLGA NS (CS-PLGA NS) showed positive zeta potential, while unmodified PLGA NS (plain-PLGA NS) were negatively charged. Decoy ODN uptake studies with Caco-2 cells using confocal laser scanning microscopy (CLSM) indicated that CS-PLGA NS were more effectively taken up by the cells than plain-PLGA NS. Decoy ODN-loaded CS-PLGA NS were able to improve the stability of ODN against DNase I or an acidic medium, such as gastric juice. Daily oral administration of CS-PLGA NS in a rat model significantly improved dextran sulfate sodium-induced diarrhea, bloody feces, shortening of colon length, and myeloperoxidase activity. Furthermore, decoy ODN-loaded CS-PLGA NS were specifically deposited and adsorbed on the inflamed mucosal tissue of the UC model rat. These results suggested that CS-PLGA NS provide an effective means of colon-specific oral decoy ODN delivery in UC., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

A new method for modifying atomic force microscope (AFM) probe with polymer was proposed to estimate the interaction between Chitosan (CS)-coated poly (lactic-co-glycolic acid) (PLGA) polymer and mucin film. In this method, the mixture of polymers and its suitable volatile dissolvent were deposited on the AFM probe tip through a micropipette controlled by micromanipulator. After being dried, the polymer film forms on the probe tip. Several evaluation experiments show that using the new method, the AFM probe tip can be smoothly and uniformly coated with PLGA and CS with approximately same curvature radius as that of probe tip less than 500 nm. As a preliminary application of the proposed method, the interaction force between PLGA/CS and mucin film in air was investigated. It was revealed that when a PLGA probe is retracting from the mucin film, a repulsive force appeared; however, after the PLGA probe was further overcoated with CS, the force became attractive if the amount of CS was enough, such as at CS concentrations of 0.2% W/V. The observed force can be interpreted by the surface hydrophilic/hydrophobic characteristics of PLGA/CS and mucin film., (Copyright 2010 Elsevier B.V. All rights reserved.)

We previously developed chitosan (CS)-modified poly (D,L-lactide-co-glycolide) (PLGA) nanospheres (NS) by an emulsion solvent diffusion method as a gene delivery system. In this study, PLGA NS were modified using polysorbate 80 (P80) to improve their cellular uptake. We investigated the cellular uptake, intracellular distribution, and transfection efficiency of P80-modified PLGA NS (P80-PLGA NS) for a plasmid DNA delivery system in A549 cells. The cellular uptake and transfection efficiency of P80-PLGA NS were greater than CS-modified PLGA NS (CS-PLGA NS). The uptake of unmodified NS and CS-PLGA NS was mediated, predominantly, by clathrin-mediated endocytosis. In contrast, a specific endocytic pathway could not be determined for the cellular uptake of P80-PLGA NS. The intracellular distributions of PLGA NS depended on their surface properties. P80-PLGA NS were not cytotoxic for A549 cells. Thus, P80-PLGA NS could be used as an effective gene delivery system; the surface properties of PLGA NS are key parameters for optimal intracellular uptake and distribution., (Copyright 2010 Elsevier B.V. All rights reserved.)

Chitosan (CS) surface-modified poly(D,L-lactide-co-glycolide) (PLGA) nanospheres (NS) for a siRNA delivery system were evaluated in vitro. siRNA-loaded PLGA NS were prepared by an emulsion solvent diffusion (ESD) method, and the physicochemical properties of NS were investigated. The level of targeted protein expression and siRNA uptake were examined in A549 cells. CS-modified PLGA NS exhibited much higher encapsulation efficiency than unmodified PLGA NS (plain-PLGA NS). CS-modified PLGA NS showed a positive zeta potential, while plain-PLGA NS were negatively charged. siRNA uptake studies by observation with confocal laser scanning microscopy (CLSM) indicated that siRNA-loaded CS-modified PLGA NS were more effectively taken up by the cells than plain-PLGA NS. The efficiencies of different siRNA preparations were compared at the level of targeted protein expression. The gene-silencing efficiency of CS-modified PLGA NS was higher and more prolonged than those of plain-PLGA NS and naked siRNA. This result correlated with the CLSM studies, which may have been due to higher cellular uptake of CS-modified PLGA NS due to electrostatic interactions. It was concluded that CS-modified PLGA NS containing siRNA could provide an effective siRNA delivery system., ((c) 2009 Elsevier B.V. All rights reserved.)

Type 2 diabetics often demonstrate normal or increased bone mineral density, yet are at increased risk for bone fracture. Furthermore, the anti-diabetic oral thiazolidinediones (PPARgamma agonists) have recently been shown to increase bone fractures. To investigate the etiology of possible structural and/or material quality defects, we have utilized a well-described mouse model of Type 2 diabetes (MKR). MKR mice exhibit muscle hypoplasia from birth with reduced mass by the pre-diabetic age of 3 weeks. A compensatory hyperplasia ensues during early (5 weeks) development; by 6-8 weeks muscle is normal in structure and function. Adult whole-bone mechanical properties were determined by 4-point bending to test susceptibility to fracture. Micro-computed tomography and cortical bone histomorphometry were utilized to assess static and dynamic indices of structure, bone formation and resorption. Osteoclastogenesis assays were performed from bone marrow-derived non-adherent cells. The 8-week and 16-week, but not 3-week, male MKR had slender (i.e., narrow relative to length) femurs that were 20% weaker (p<0.05) relative to WT control femurs. Tissue-level mineral density was not affected. Impaired periosteal expansion during early diabetes resulted from 250% more, and 40% less of the cortical bone surface undergoing resorption and formation, respectively (p<0.05). Greater resorption persisted in adult MKR on both periosteal and endosteal surfaces. Differences were not limited to cortical bone as the distal femur metaphysis of 16 week MKR contained less trabecular bone and trabecular separation was greater than in WT by 60% (p<0.05). At all ages, MKR marrow-derived cultures demonstrated the ability for enhanced osteoclast differentiation in response to M-CSF and RANK-L. Taken together, the MKR mouse model suggests that skeletal fragility in Type 2 diabetes may arise from reduced transverse bone accrual and increased osteoclastogenesis during growth that is accelerated by the diabetic/hyperinsulinemic milieu. Further, these results emphasize the importance of evaluating diabetic bone based on morphology in addition to bone mass.

The aim of the present study was to determine whether the antiobesity effects of tea catechins (TCs) are associated with the expression of uncoupling protein 1 (UCP1) in brown adipose tissue (BAT). Male Sprague-Dawley rats were fed a high-fat (HF; 35% fat) diet for 5 weeks, then divided into four groups and fed an HF, HF with 0.5% TC (HFTC), normal-fat (NF; 5% fat) or NF with 0.5% TC (NFTC) diet for 8 weeks. At the end of the experimental period, perirenal and epididymal white adipose tissues (WATs) and interscapular BAT were isolated. The NFTC group had significantly lower perirenal WAT weights than the NF group (NF: 12.7+/-0.53 g; NFTC: 10.2+/-0.43 g; P<.01), but the HF and HFTC groups did not differ significantly. TC intake had no effects on epididymal WAT weights. The NFTC and HFTC groups had significantly lower BAT weights than the NF and HF groups, respectively. The NFTC group had significantly higher UCP1 mRNA levels in BAT than the NF group (NF: 0.35+/-0.02; NFTC: 0.60+/-0.11; P<.05), but the HF and HFTC groups did not differ significantly. Thus, TC intake in the context of the NF diet reduced perirenal WAT weight and up-regulated UCP1 mRNA expression in BAT. These results suggest that the suppressive effect of TC on body fat accumulation is associated with UCP1 expression in BAT.

Objective: The aim of this study is to clarify the etiology of first-bite syndrome., Methods: A total of 29 consecutive patients treated with a surgical resection for parapharyngeal space tumor between April 1999 and April 2005 were retrospectively reviewed., Results: Nine cases of first-bite syndrome were developed. Eight of those patients had undergone surgical ablation of the cervical sympathetic chain and/or external carotid artery. In two patients who underwent ablation of the sympathetic chain but in whom first-bite syndrome did not develop, the superior cervical ganglion was probably preserved., Conclusion: The present findings strongly indicate that loss of sympathetic innervation to the parotid gland from the superior cervical ganglion causes first-bite syndrome. They also suggest that some residual or autonomous activity of the superior cervical ganglion could prevent development of first-bite syndrome even if the lower sympathetic trunk was ablated.

Novel 2-deoxo-2-phenyl-5-deazaflavins and 2-deoxo-2-phenylflavin-5-oxides were prepared as a new class of antitumor agents and showed significant antitumor activities against NCI-H 460, HCT 116, A 431, CCRF-HSB-2, andKB cell lines. In vivo investigation, 2-deoxo-10-methyl-2-phenyl-5-deazaflavin exhibited the effective antitumor activity against A 431 human adenocarcinoma cells transplanted subcutaneously into nude mouse. Furthermore, AutoDock study has been done by binding of the flavin analogs into PTK pp60(c-src), where a good correlation between their IC(50) and AutoDock binding free energy was exhibited. In particular, 2-deoxo-2-phenylflavin-5-oxides exhibited the highest potential binding affinity within the binding pocket of PTK.

Mammalian endogenous circadian rhythms are entrained to the environmental light-dark (LD) cycle. Although the circadian rhythms of core body temperature (Tb) and spontaneous locomotor activity (LA) are well synchronized under stable LD conditions, it is thought that these two parameters are regulated by distinct mechanisms. The purpose of the present study was to examine the adaptability of these two rhythms to an abrupt change in the environmental light phase. Tb and LA were simultaneously recorded in individual mice kept under 12:12-h LD cycle conditions before and after an 8-h photic phase advance. The onset of LA required 8 days to reentrain to the new LD cycle, whereas 6 days were required for reentrainment of the acrophase of Tb. Resting Tb, i.e., the Tb level independent of LA, was extracted from the same data source. The resting Tb level exhibited a robust daily rhythm with a difference of 1.0 degrees C between LD phases. After the photic phase advance, the resting Tb rapidly reached a stable level within 4 days, whereas the uncorrected Tb required 6 days for reentrainment. Based on these findings, we revealed that, independent of LA, the adaptability of the Tb rhythm to a new light cycle is half as rapid as that of LA. These results therefore suggest that the circadian rhythms of Tb and LA are intrinsically regulated by different pacemaker or effector mechanisms.

Pruritus is a common, distressing and difficult to manage complication of many autoimmune diseases. A suitable animal model of autoimmune disease associated pruritus would contribute to a better understanding of the pathophysiology of this symptom and lead to the development of safe and effective antipruritic agents. We noticed spontaneous scratching behavior in aged MRL/lpr mice, a model of autoimmune disease. This scratching behavior was observed in a specific pathogen-free environment and was more frequent in female mice. In contrast to animal models of dermatitis; NC/Nga mice, the serum IgE and IgG1 levels in MRL/lpr mice were not elevated. These features indicate that this scratching behavior is similar to human autoimmune disease associated pruritus. The antipruritic effects of an antihistamine (chlorpheniramine), an opioid receptor antagonist (naltrexone), and a novel kappa-opioid receptor agonist (nalfurafine hydrochloride [TRK-820]) were evaluated. The frequency of scratching was not reduced by oral administration of chlorpheniramine, suggesting that the behavior is antihistamine-resistant. The oral administration of nalfurafine and subcutaneously administered naltrexone inhibited the scratching behavior without causing gross behavioral changes. In conclusion, MRL/lpr mice scratching behavior is a suitable model of pruritus that occurs in autoimmune diseases, and nalfurafine was shown to be efficacious against this behavior suggesting that it may be beneficial in patients with autoimmune disease associated pruritus.

In Japan, mumps vaccination is optional and the mumps were increased accompanied with prevalent waves according to the Infections Agents Surveillance Report (IASR). The aim of this study was to clarify that there was relevance for increase of mumps epidemic and mumps deafness. The Acute Profound Deafness Research Committee of the Japanese Ministry of Health and Welfare (reorganized to the Ministry of Health, Labour and Welfare in 2001) conducted a nationwide epidemiological survey to determine the number of patients treated for mumps deafness in 1987, 1993 and 2001. Based on its findings, the annual numbers of mumps deafness cases was estimated to be 300 in 1987, 400 in 1993 and 650 in 2001, which correlated with the overall incidence of mumps in those years. Because the majority of cases exhibited severe or profound sensorineural hearing loss that usually did not recover, rapid improvement of mumps vaccine coverage is strongly recommended.

5-Fluorouracil (5-FU) in combination with leucovorin (LV) is nowadays the standard treatment in colon cancer and would be a candidate to be delivered orally to the colon. Eudragit P-4135F or Eudragit RS100 were used separately to prepare microspheres by an oil/oil emulsification process trapping 5-FU and LV simultaneously. Scanning electron microscopy permitted a structural analysis, process parameters were analyzed and drug loading and release profiles were recorded. Particle size varied between 123 (RS100) and 146 microm (P-4135F). Generally, higher encapsulation rates were found with RS100 (5-FU, 60.3+/-9.7%; LV, 81.4+/-8.6%) compared to P-4135F (5-FU, 48.3+/-2.0%; LV, 55.4+/-2.7%). Microparticles made from Eudragit RS100 released the incorporated drug combination within 8 h not exhibiting general differences between the kinetics of both drugs. P-4135F was found to maintain the undesired 5-FU release at pH 6.8 lower than 25% within 4 h while at pH 7.4, a nearly immediate release (within 15 min) was observed. Although the release was similar at pH 7.4, at pH 6.8 LV showed a distinct initial drug loss of about 60% and a complete release within 2 h. SEM analyses revealed a substantial presence of LV crystals on the particle surface provoking a distinct burst effect of LV. These observations were concluded to be related to the high lipophilicity of P-4135F provoking a separation between P-4135F and LV during the preparation process.

FK888 is a candidate selective NK1 receptor antagonist, and it exhibits poor absorption from the gastrointestinal tract in healthy volunteers. In a previous study, the optimized dry powder inhaler (DPI) formulation with carrier lactose using the Spinhaler was developed, although the maximum dose per capsule was only 5mg because the fine particle fraction (FPF) was reduced at doses over 5mg. The objective of this study was to develop an optimized DPI formulation for higher doses, such as 40 mg, with proportional systemic absorption. The Spinhaler and E-haler were used as the inhalation devices, and the in vitro deposition was evaluated using a multistage cascade impactor at different flow rates (28.3 and 60 l/min). When hydroxypropyl methylcellulose (HPMC) capsules were used as the container, and spherical soft agglomerates of fine FK888 particles (soft pellets) and the E-haler were used, the fraction of particles emitted from the inhalation system (Em) was significantly improved, to over 80% of the nominal dose, and no significant difference was found between the airflow rates (84.3+/-2.3% for 28.3 l/min, 88.1+/-3.6% for 60 l/min). It was also found that the E-haler was an extremely suitable device for obtaining the higher respirable particle percentage of emitted particles (RP) in the 40 mg formulation with the soft pellets contained in HPMC capsules (35.0+/-1.8% for 28.3 l/min and 42.5+/-3.5% for 60 l/min), compared with the Spinhaler (13.8+/-3.0% for 28.3 l/min and 28.9+/-1.0% for 60 l/min). Using the formulations with the E-haler, proportional systemic absorption was achieved up to 40 mg FK888 in healthy volunteers (62.91+/-27.58, 103.70+/-40.19 and 254.79+/-85.01 ngh/ml as AUCs for 10, 20 and 40 mg FK888, respectively; R(2)=0.9641). It is also expected that the E-haler will act as an efficient device when a higher dose, such as 40 mg, is required in clinical situations.

Recently, tacrolimus was shown to be effective in mitigating inflammatory bowel disease (IBD). In the treatment of IBD, oral drug delivery using pH-dependent polymers is one of the most successful therapeutic strategies. Eudragit P-4135F, a pH-sensitive polymer for colonic delivery was used to prepare tacrolimus microparticles using an oil/oil emulsification or an oil/water emulsification technique combined with a solvent extraction or evaporation step. Although the pH-dependent drug release was similar for all types of microspheres, it was generally found that encapsulation rates of oil/water systems (extraction 38.8 +/- 9.4%; evaporation 56.3 +/- 1.9%) were superior to the oil/oil emulsification (4.8 +/- 0.4%). Eudragit P-4135F was found to limit drug leakage at pH 6.8 to levels lower than 10% within 6 h. At pH 7.4, almost immediate release (within 30 min) was observed. From differential scanning calorimetry and X-ray analyses, the drug inside the polymeric microsphere matrix was concluded to be present in a molecular dispersion. Generally, all formulations proved their applicability in vitro as a promising device for pH-dependent colonic delivery of tacrolimus, however, the oil/water technique was found to be superior to the oil/oil approach and among them solvent evaporation seemed to be more advisable, due to the higher encapsulation rate., (Copyright 2004 Elsevier B.V.)

Hollow microspheres (microballoons) floatable in JPX III No.1 solution were developed as a dosage form characterized by excellent buoyant properties in the stomach. Microballoons were prepared by the emulsion solvent diffusion method utilizing enteric acrylic polymers codissolved with drug in a mixture of dichloromethane and ethanol. The release properties of five different drugs exhibiting distinct water solubilities (aspirin, salicylic acid, ethoxybenzamide, indomethacin and riboflavin) entrapped within microballoons were investigated. Buoyancy of the microballoons decreased with increasing drug release rate. In the case of aspirin, salicylic acid and ethoxybenzamide, the drug release profiles of microballoons proved a linear relationships by Higuchi plotting. However, indomethacin and riboflavin release profiles did not follow the Higuchi equation. When the loading amount of riboflavin was higher than the solubility in the mixture of dichloromethane and ethanol, the drug release profiles of the microballoons displayed an initial burst release. The insoluble riboflavin in the mixture of dichloromethane and ethanol adsorbed on to the microballoon surface in the crystal state. Such riboflavin crystals were released preferentially at the initial stage of the release test, which was attributable to the initial burst. In addition, by incorporating a polymer such as hydroxypropylmethylcellulose within the shell of microballoons, the release rate of riboflavin from the microballoons could be controlled while maintaining high buoyancy.

Sodium guaiazulene sulfonate (GAS-Na), which has an anti-inflammatory effect, is an unstable compound, which is gradually decomposed in the solid state at room temperature. In fact, when heated (40 degrees C 6% RH), GAS-Na decomposes almost completely within 1 week. It was found that a kneaded mixture of GAS-Na and cornstarch (weight ratio; 1:250) for tableting with water is stable. So, during production, GAS-Na could be stabilized using water. Four kinds of tablet were prepared in different ways: direct tableting, tableting via screw granulation, tableting via fluidized bed granulation, and tableting via twin-screw extrusion. The stability of GAS-Na in these tablets was compared. The tablet prepared using screw granulation, during which 30% water was added to the material, was the most stable. It was, however, shown that reducing the water content to 12.5%, when screw granulation was conducted, made the GAS-Na less stable. Also, when a twin-screw extruder with kneading paddle elements in the screws was used even with lower water content of 12.5%, the stability of GAS-Na improved. In addition, when the kneading paddle elements were detached from the screws and only the feed screw elements were operated, GAS-Na lost its stability. These results show that the kneading paddle elements play a role in uniformly dispersing a small amount of water into the powder and stabilizing GAS-Na. It was found that the water presence was a very important factor with respect to the decomposition of GAS-Na, irrespective of the crystallinity. Furthermore, a twin-screw extruder with kneading paddle elements is useful for uniformly dispersing water to prepare stable formulations of GAS-Na.

FK224 is a cyclopeptide drug with poor oral absorption due to proteolysis in the gastrointestinal tract. The objectives of this study were to investigate the absorption of FK224 from the lung in healthy volunteers, and compare the pharmacokinetic profiles of FK224 after inhalation from a pressurized metered dose inhaler (pMDI) and dry powder inhaler (DPI). The pMDI (Suspension type, 1 mg as FK224/puff) and DPI (4 mg and 10 mg as FK224/capsule, using Spinhaler as the device) were developed by formulating the same micronized particles of FK224 which were premixed with beta-cyclodextrin (beta-CyD) to improve the solubility of FK224. In the case of pMDI, 1, 4 or 8 mg was inhaled by the corresponding number of puffs with the pMDI. In addition, the in vitro drug delivery characteristics of the inhalers were evaluated using a multistage liquid impinger. In both inhalers, it was observed that FK224 could be absorbed into the systemic circulation from the lungs of the healthy volunteers, and the AUC and C(max) were proportionally increased depending on the emitted dose after inhalation. However, the pharmacokinetic (PK) parameters for DPI were significantly higher than that of pMDI, in spite of usage of the same fine particles for the formulations in both inhalers. Based on the distribution from the in vitro examination, the fine particle dose, which is defined as the dose region delivered as particles <3.8 microm, was calculated from the emitted dose inhaled by the healthy volunteers. It was found that the PK parameters for both inhalers were proportionally increased depending on the predicted fine particle dose regardless of the type of inhaler. This suggests that the absorption from the lung is influenced by the fine particle dose. We concluded that DPI is a suitable inhaler for FK224, and the alveolus, which is generally known as the site of action of the fine particles, is a possible absorptive site for FK224.

The purpose of the present study was to determine whether dehydroepiandrosterone (DHEA) affects de novo fatty acid synthesis, oleic acid formation, fatty acid oxidation, and very low density lipoprotein (VLDL) secretion, in relation to the accumulation of lipid containing oleic acid, in rat liver. The rates of hepatic de novo synthesis of both fatty acid and monounsaturated fatty acid, determined by incorporation of 3H from 3H(2)O into fatty acid, were increased markedly when rats were fed a diet containing 0.5% (w/w) DHEA for 14 days. The treatment of rats with DHEA also enhanced the conversion of [14C]stearic acid into oleic acid in the liver in vivo. DHEA did not suppress fatty acid degradation in the liver. Namely, mitochondrial palmitic acid oxidation in liver homogenates and isolated hepatocytes was increased approximately 1.9- and 5-fold, respectively, in DHEA-treated rats. Peroxisomal palmitic acid oxidation in isolated hepatocytes from rats treated with DHEA, however, was not significantly different from that of the control, despite the fact that peroxisomal degradation of palmitic acid in the liver homogenates was increased markedly. The rate of hepatic VLDL secretion in DHEA-treated rats was decreased markedly. These results indicate that the elevation of the hepatic fatty acid content, especially oleic acid, by DHEA feeding is due to an increase in both de novo fatty acid synthesis and the formation of oleic acid and to a decrease in the rate of hepatic VLDL secretion. Mitochondrial and peroxisomal fatty acid degradation does not appear to play a significant role in the accumulation of hepatic lipids.

Hollow microspheres (microballoons) floatable on JPXIII No.1 solution were developed as a dosage form capable of floating in the stomach. Hollow microspheres were prepared by the emulsion solvent diffusion method using enteric acrylic polymers with drug in a mixture of dichloromethane and ethanol. It was found that preparation temperature determined the formation of cavity inside the microsphere and the surface smoothness, determining the floatability and the drug release rate of the microballoon. The correlation between the buoyancy of microballoons and their physical properties, e.g. apparent density and roundness of microballoons were elucidated. The drug loading efficiency of microballoons with various types of drug was investigated and correlated to the distribution coefficient of drug between dichloromethane and water. The optimum loading amount of riboflavin in the microballoon was found to impart ideal floatable properties to the microballoons. On the other hand, little entrapment was observed for aspirin due to the low distribution coefficient; however, entrapment improved to some extent upon reduction of the pH of the process.

FK224 is a cyclopeptide drug with a low aqueous solubility. Following oral administration to rats, poor absorption was observed due to proteolysis in the gastrointestinal tract. The objective of this study was to investigate the effect of the pulmonary route on the systemic absorption of FK224 in comparison with other administration routes, and to determine the bioavailability (BA) of FK224 following pulmonary administration in rats using various dosage forms. From absorption studies on the Polyethylene Glycol 400 solution given by various routes (intranasal, subcutaneous, intratracheal and intravenous as reference), it was shown that pulmonary administration was a potentially attractive route for FK224. In the pulmonary absorption studies, after administration of the aqueous suspension, the BA was reduced to 2.7% compared with 16.8% for the solution. However, beta-cyclodextrin (beta-CyD) was found to be an effective additive as far as improving the solubility of FK224 was concerned. The BA of the aqueous suspension containing beta-CyD was increased to 19.2%. Pressurized metered dose inhalers were prepared by formulating beta-CyD with various molar ratios of 1:0, 1:1 and 1:7 (FK224/beta-CyD), and the resulting BAs were 4.3%, 29.0% and 91.2%, respectively. It was observed that both the C(max) and AUC of FK224 were increased as the amount of beta-CyD increased. The plasma profiles showed sustained absorption. In conclusion, we have seen that the lung is a suitable route for absorption of FK224, and beta-CyD is an extremely effective additive as far as improving the pulmonary absorption of FK224 is concerned. beta-CyD or derivatives with various degrees of aqueous solubility are potential drug carriers for controlling pulmonary absorption., (Copyright 2002 Elsevier Science B.V.)

The mechanism by which p-chlorophenoxyisobutyric acid (clofibric acid) induces striking changes in the proportion of the molecular species of phosphatidylcholine (PC) in rat liver was studied. Treatment of rats with clofibric acid strikingly increased the content of 1-palmitoyl-2-oleoyl (16:0-18:1) PC, but decreased the contents of 1-palmitoyl-2-docosahexaenoyl (16:0-22:6), 1-stearoyl-2-arachidonoyl (18:0-20:4), and 1-stearoyl-2-linoleoyl (18:0-18:2) PC; the drug did not change the content of 1-palmitoyl-2-arachidonoyl (16:0-20:4) PC. The mechanism underlying these changes has been investigated with regard to the in vivo formation of the molecular species of PC by: (i) de novo synthesis, (ii) reacylation, and (iii) methylation of phosphatidylethanolamine (PE). We found that (i) the incorporation of [3H]glycerol, which was injected intravenously, into 16:0-18:1 diacylglycerol (DG) and 16:0-18:1 PC was increased markedly by clofibric acid feeding without changing the substrate specificity of CDP-choline:DG cholinephosphotransferase, (ii) the in vivo formation of 16:0-18:1 and 16:0-20:4 PC from 1-16:0-[3H]glycerophosphocholine (GPC), which was injected intraportally, was increased markedly by clofibric acid feeding, and (iii) the incorporation of [14C]ethanolamine, which was injected intravenously into 16:0-22:6, 18:0-22:6, and 18:0-20:4 PC, was decreased by clofibric acid feeding; the extent of the decrease in 16:0-20:4 PC was less than that of 18:0-20:4 PC. It was concluded, therefore, that (i) clofibric acid selectively increased the content and proportion of 16:0-18:1 PC by enhancing both the CDP-choline pathway and the remodeling of the pre-existing PC molecule, and (ii) the drug kept the content of 16:0-20:4 PC unchanged by stimulating the remodeling of the pre-existing PC molecule, whereas the formation of other more long chain, polyunsaturated molecular species, such as 16:0-22:6, 18:0-22:6, and 18:0-20:4, was decreased owing to the suppression of PE methylation.

The effects of dehydroepiandrosterone (DHEA) on the acyl composition of lipids in rat liver were studied. The content of oleic acid (18:1) in hepatic lipids was increased markedly by feeding rats a diet containing 0.5% (w/w) DHEA for 14 days. Treatment of rats with DHEA caused an increase in the activity of the terminal desaturase of the stearoyl-CoA desaturation system, without changing either the activity of NADH-cytochrome b5 reductase or the microsomal content of cytochrome b5. Among the changes observed in hepatic lipids, the increase in 18:1 content in phosphatidylcholine (PtdCho) was the most prominent; an approximately 2.5-fold increase in the proportion of 18:1 was induced at position 2, but not at position 1, by DHEA. This selective elevation of 18:1 at position 2 of PtdCho seems to be produced by the concerted actions of the induced 1-acylglycerophosphocholine (1-acyl-GPC) acyltransferase and the induced stearoyl-CoA desaturase. The content of 18:1 in serum lipids was unchanged by DHEA treatment, suggesting that secretion of lipids containing 18:1 into the circulation was not affected by DHEA. These results suggest that the elevation of hepatic content of 18:1 caused by DHEA treatment is mainly due to the induction of stearoyl-CoA desaturase.

The purpose of this study was to evaluate the functions of a modified polyvinyl alcohol (PVA-R), which has a hydrophobic moiety, as a coating material for liposomes to be loaded with the anticancer drug, doxorubicin. The size controlled liposomes (egg phosphatidylcholine: cholesterol=1:1 molar ratio) were prepared by the hydration method followed by extrusion. Drug encapsulation and surface modification with polymers (PVA and PVA-R) were carried out simultaneously using a modified pH gradient method. The existence of a thick polymer layer on the surface of the liposomes was confirmed by an increase in particle size and the amount of polymer on the liposomal surface, especially for the PVA-R-coated liposomes. The effects of polymer coating on the behavior of the liposomes in vivo were evaluated by measuring the circulation time and biodistribution of the drug after i.v. administration of the liposomal drug in rats. The PVA-R-coated liposomes showed a more prolonged circulating time for the drug with less uptake by the reticuloendothelial system after i.v. administration in rats, compared with non-coated liposomes. These results confirm that polymer possessing a hydrophobic anchor at its end, like PVA-R, is a suitable material for modifying the surface of doxorubicin-loaded liposomes to improve their stability in the circulating blood.

Metabolic changes induced by 2,2'-(decamethylenedithio)diethanol (tiadenol) and di-(2-ethylhexyl)phthalate (DEHP) in the biosynthesis of phosphatidylcholine (PtdCho) and phosphatidylethanolamine (PtdEtn) in rat liver were compared with changes induced by p-chlorophenoxyisobutyric acid (clofibric acid). Treatment of rats with either tiadenol or DEHP increased the hepatic contents of PtdCho and PtdEtn, as was observed with clofibric acid treatment. The administration of tiadenol, DEHP, or clofibric acid slightly, but significantly, increased, in common, the activity of CTP:phosphocholine cytidylyltransferase, a key enzyme for the synthesis de novo of PtdCho, and suppressed the activity of PtdEtn N-methyltransferase. With regard to the enzymes involved in the synthesis of PtdEtn, the three peroxisome proliferators enhanced the activity of phosphatidylserine (PtdSer) decarboxylase and markedly decreased the activity of CTP:phosphoethanolamine cytidylyltransferase. Treatment of rats with the three compounds markedly increased, in common, the content and the proportion of the molecular species of PtdCho containing oleic acid (18:1), but considerably decreased the proportion of the molecular species of PtdCho containing linoleic acid (18:2) in the liver, resulting in a striking decrease in the concentration of the molecular species of PtdCho containing 18:2 in the serum. The present study suggests that the administration of peroxisome proliferators to rats increases the contents of hepatic PtdCho and PtdEtn for hepatomegaly and proliferation of organelles by the same mechanism, irrespective of their chemical structures.

A combined method of immunoaffinity extraction with high-performance liquid chromatography has been developed for the enantioselective determination of bufuralol and its metabolites in human plasma. The antibodies having high affinity toward the asymmetric center at the C-1 position of bufuralol and its 1'-oxidized metabolites and low affinity to their antipodes were elicited by immunization of rabbits with immunogens, (1R)- and (1S)-1'-oxobufuralol O-carboxymethyloxime-bovine serum albumin conjugates, respectively. 0.5 ml Of the immunoaffinity adsorbent (7.6 mg.ml-1 for anti-(1S)-antibody and 28.8 mg.ml-1 for anti-(1R)-antibody) prepared by immobilization of an antibody was capable of retaining up to 1 microgram of (R)- and (S)-bufuralol and up to 500 ng of other metabolites. The adsorbates were recovered quantitatively by elution with methanol-10 mM ammonium acetate buffer (pH 5) (95:5, v/v) without any interfering peaks on the high-performance liquid chromatogram. The proposed method was evaluated to be useful for the simultaneous determination of optically active bufuralol and its metabolite in plasma with acceptable recovery and precision.

Novel emulsion solvent diffusion methods in water or oil were developed to prepare the peptide (TRH and elcatonin)-loaded PLGA nanospheres, via the coprecipitation of drug and polymer in the emulsion droplets induced by the diffusion of solvent. The PLGA nanospheres prepared by the emulsion solvent diffusion method in water were unimodally dispersed particles with an average diameter of about 250 nm, whereas those prepared by the emulsion solvent diffusion method in oil and the phase separation method exhibited bimodal distribution with average diameter of about 700 and 800 nm, respectively. The content and recovery of the peptide and nanospheres prepared by the emulsion solvent diffusion method in oil were superior to those of nanospheres prepared using other methods. The drug release properties from nanospheres depended strongly on their preparation processes. The nanospheres prepared by the emulsion solvent diffusion method in oil, released the drug continuously over 14 days, the rate of which was determined by the diffusion of drug in the rigid matrix structure of the nanosphere. The drug release behavior of nanospheres prepared by the phase separation method exhibited a triphasic release pattern with an initial burst and an induction period followed by the diffusion of drug through the walls of the reservoir type nanospheres, which were produced via the phase separation of polymer deposited around the emulsion droplets of the drug.

A set of the title compounds having different substituents (R1, R2) on their phenyl groups was synthesized to find sigma receptor binding affinity. Among the compounds, 2b (R1 = R2 = Cl) has the most potent sigma 1-binding activity, while 2a (R1 = R2 = H, SA4503) was most selective to sigma 1 over sigma 2 receptor. The crystal structures of 2a and 2b were shown, by X-ray crystallography, to be similar except for the one torsional angle of their propylene parts. Quantitative structure-activity relationship study suggested the affinity of the compounds to the sigma 1 receptor was dependent on the electronic feature, Swain-Lupton's R or Sz that was derived by molecular orbital method, of R1 and R2.

The binding profiles of SA4503 (1-(3,4-dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine dihydrochloride), a novel sigma receptor ligand, to sigma 1 and sigma 2 receptor subtypes in guinea pig and rat brain membranes were evaluated. SA4503 showed a high affinity for the sigma 1 receptor subtype labeled by (+)-[3H]pentazocine (IC50 = 17.4 +/- 1.9 nM), while it had about 100-fold less affinity for the sigma 2 receptor subtype labeled by [3H]1,3-di(2-tolyl)guanidine ([3H]DTG) in the presence of 200 nM (+)-pentazocine. SA4503 showed little affinity for 36 other receptors, ion channels and second messenger systems. The inhibition curves of SA4503 for (+)-[3H]pentazocine binding were shifted to the right in the presence of guanosine 5'-o-(3-thiotriphosphate) (GTP gamma S), as similar to those of (+)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine ((+)-3-PPP) and (+)-pentazocine, sigma 1 receptor agonists. SA4503 significantly increased the KD value, but did not affect the Bmax value for specific (+)-[3H]pentazocine binding. These results indicated that SA4503 is a potent and selective agonist for the sigma 1 receptor subtype in the brain. In addition, SA4503 inhibited specific (+)-[3H]pentazocine binding in a competitive manner.

1. The ability of VA-045 to improve aged-related impairment on electroencephalograph (EEG), caudate spindle, performance on a passive avoidance task and cerebral blood flow (CBF) were evaluated in rats. 2. The cortical EEG of the aged rats showed a higher incidence of spontaneous spindle burst (SSB) than seen in young rats. VA-045 decreased the incidence of SSB in aged rats. In contrast, vinpocetine increased the incidence of SSB in aged rats. 3. Electrical stimulation of the striatum in aged rats lead to a higher incidence of neocortical high voltage spindle (CS) than seen in young rats. In young rats, VA-045 had no effect on the CS, whereas an age-related increase in CS was blocked by VA-045, but was enhanced by vinpocetine. 4. There were no differences in the cortical EEG arousal response elicited by stimulation of the reticular formation of the brain stem in rats of all ages. VA-045 and vinpocetine had no effect on the cortical EEG arousal response in both young and aged rats. 5. VA-045, but not vinpocetine, attenuated the age-related decreased step through latency (STL) on a passive avoidance task. VA-045 and vinpocetine did not enhance the acquisition of learning behavior in a passive avoidance task in young rats. 6. VA-045 increased the cerebral blood flow (CBF) in both young and aged rats and the potency in aged rats was greater than that in young rats. Vinpocetine had no effect on CBF in either young or aged rats. 7. The pharmacological effects of VA-045 on age-related neuronal dysfunction are discussed.

Nuclear magnetic resonance (NMR) studies were carried out for the title compound ((+/-)-3,4-dihydro-2-[5-methoxy-2- [3-[N-methyl-N-[2-(3,4-methylenedioxy)phenoxy]ethyl]amino] propoxy]phenyl]-4-methyl-3-oxo-2H-1,4-benzothiazine) (1a; R = H) and its 2-substituted analogs (1b; R = OCH3, 1c; R = SCH3, 1d; R = CH3, 1e; R = i-C3H7) which had Ca2+ antagonistic activities. Conformational analysis using the model compounds of the 2-phenylbenzothiazine (2-PBT) part of 1 by semiempirical molecular orbital method agreed with the NMR behavior. Two local minimum conformations, having different rotational angles (theta 1) of the 2-phenyl ring, were suggested for biologically active 1a-1d. The molar fractional ratios, including the conformations within a particular theta 1 range that contained each global minimum conformation, were found to correlate well with the activities. In the same theta 1 range, any stable conformation was not indicated for non-active compound 1e. From these results, the active conformation for the 2-PBT part of 1 was suggested to be similar to the global minimum conformation indicated for the most potent 1a.

Alterations induced by p-chlorophenoxyisobutyric acid (clofibric acid) in the composition of phosphatidylcholine and cholesterol esters in serum and their influence on the composition of phosphatidylcholine in the kidney were studied. Rats of different ages responded differently to the drug in terms of the levels of arachidonic acid (20:4) and linoleic acid (18:2) in the phosphatidylcholine and cholesterol esters in the serum. Administration of clofibric acid to 26-week-old rats for 2 weeks caused a marked decreased in the relative level of 20:4 in phosphatidylcholine and cholesterol esters in serum, whereas similar treatment of 6-week-old rats resulted in a reduction of 18:2 and, to a lesser extent, of 20:4 in serum lipids. The decrease in phosphatidylcholine that contained 20:4 in the serum of old rats was mainly due to a decrease in the concentration of stearoyl-arachidonoyl (18:0-20:4) species. The decrease in cholesterol arachidonate in serum caused by the treatment of old rats with clofibric acid seemed to be due to a reduction in the relative level of serum phosphatidylcholine containing 20:4. The marked reduction in serum lipids that contained 20:4 caused a decrease in the relative level of 20:4 in renal phospholipids, in particular, a decrease in the proportion of palmitoyl-arachidonoyl (16:0-20:4) and 18:0-20:4 phosphatidylcholine.

Response of rat kidney to the challenges by perfluorooctanoic acid (PFOA) was studied using microsomal 1-acyglycerophosphocholine (1-acyl-GPC) acyltransferase as a parameter. Marked induction of the enzyme was brought about in kidney of male rats, whereas the induction in kidney of female rats was far less pronounced. The sex-related difference in the response of kidney to PFOA was much more marked than those seen with p-chlorophenoxyisobutyric acid (clofibric acid) or 2,2'-(decamethy-lenedithio)diethanol (tiadenol). Hormonal manipulations revealed that the sex-related difference in the response of kidney to PFOA was strongly dependent on the state of gonadal hormones of rats. Even after a prolonged administration of PFOA for up to 26 weeks, this sex-related difference was still evident. Induction of peroxisomal beta-oxidation was brought about concurrently with microsomal 1-acyl-GPC acyltransferase and a high correlation was confirmed between the inductions of these two parameters.

Changes in the levels of rapid turnover proteins (RTP) during total parenteral nutrition (TPN) were compared with changes in other nutritional measures in 40 adult patients with benign diseases who were considered to have moderate to severe protein-energy malnutrition on the basis of anthropometric measurements. Measures such as weight for height, arm muscle circumference, triceps skinfold thickness, and serum albumin showed significant increases by the third to fourth week of TPN, but significant increases in RTP were already observed during the first week. The per cent changes in RTP levels compared with the levels before TPN were significantly greater than the changes in the other measures. The RTP are sensitive indices of protein nutritional status and should be efficacious for the evaluation of nutritional therapy.

A real-time Automated Computerized Rat Eater Meter was developed by modifying commercially available metabolic cages. Food access via a feeding tunnel was monitored by photocells. Food consumption was measured by an electronic scale. The signals thus generated were processed by a computer. This allowed us to continuously measure the spontaneous feeding behavior of free-feeding nondeprived Fischer rats for a sum total of 35 study days. Based on our data, we defined a meal as an episode of food consumption preceded and followed by at least 5 minutes of no feeding. Fischer rats showed periodic nychthemeral eating behavior. Food consumption, number of meals, meal sniffs, intermeal sniffs, and, consequently, eating activity were greater during the dark cycle than the light cycle. Meal duration, meal size, and thus food consumption rates remained constant throughout both cycles. Our modification of commercially available metabolic cages provides unique data for continuously monitoring rat feeding patterns over prolonged periods of time.

Long-term effects of rho-chlorophenoxyisobutyric acid (clofibric acid) on inductions of peroxisomal beta-oxidation, fatty acid-binding protein and cytosolic acyl-CoA hydrolases in rat liver were studied. Male rats were fed clofibric acid at a dietary concentration of 0.25% for 22 weeks. The induction of peroxisomal beta-oxidation activity lasted throughout the long-term treatment of rats, the activity being a half that of rats treated with clofibric acid for 2 weeks. cytosolic long-chain acyl-CoA hydrolase I and II were both induced by the long-term and the short-term treatment of age-matched rats with clofibric acid, although the ability to induce hydrolase I decreased greatly by aging of rats. There was little difference in the inducing effect on fatty acid-binding protein between the long-term treatment and the short-term treatment. These results suggest that the inductions of peroxisomal beta-oxidation, fatty acid-binding protein and two cytosolic long-chain acyl-CoA hydrolases are essential responses of rats to clofibric acid (but not the brief events which occur in only the first stage of the continuous treatment with clofibric acid).

The effects of feeding 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) on the level of peroxisomal enzymes in rat liver were studied. The concentration of triglyceride in serum was decreased and the activity of cyanide-insensitive palmitoyl-CoA oxidation, catalase and carnitine acetyltransferase increased. However, the extent of the increase in the activity of these enzymes by treatment with 2,4-D was less pronounced than that by 2,4,5-T treatment. The administration of 2,4-D or 2,4,5-T increased the concentration of polypeptide with a mol. wt of 80,000 in the light mitochondrial fractions of the liver from the rats.

Long-term effects of p-chlorophenoxyisobutyric acid (clofibric acid) on inductions of stearoyl-CoA desaturase and 1-acylglycerophosphorylcholine (1-acyl-GPC) acyltransferase, and on changes in fatty acyl composition of microsomal lipid in rat liver were studied. Male rats were fed clofibric acid at a dietary concentration of 0.25% for 2 or 22 weeks. Inductions of stearoyl-CoA desaturase and 1-acyl-GPC acyltransferase lasted throughout the long-term treatment and were the same as those of either young or aged rats which were treated with clofibric acid for 2 weeks. The long-term treatment of rats with clofibric acid scarcely affected components of stearoyl-CoA desaturation system other than terminal desaturase. In accordance with the induction of stearoyl-CoA desaturase, the increase in the proportion of octadecenoic acid in hepatic lipid lasted throughout the 22-week treatment. In the case of both of the long-term treatment and the short-term treatment of rats, the increase in the proportion of octadecenoic acid in microsomal phosphatidylcholine was due to the marked increase in the proportion of octadecenoic acid in position 2, but not position 1, of phosphatidylcholine. These changes in fatty acyl composition of phosphatidylcholine were not due to the alteration of the content of phosphatidylcholine in liver.